Effect of taurine on acinar cell apoptosis and pancreatic fibrosis in dibutyltin dichloride-induced chronic pancreatitis.

The relationship between pancreatic fibrosis and apoptosis of pancreatic acinar cells has not been fully elucidated. We reported that taurine had an anti-fibrotic effect in a dibutyltin dichloride (DBTC)-chronic pancreatitis model. However, the effect of taurine on apoptosis of pancreatic acinar cells is still unclear. Therefore, we examined apoptosis in DBTC-chronic pancreatitis and in the AR42J pancreatic acinar cell line with/without taurine. Pancreatic fibrosis was induced by a single administration of DBTC. Rats were fed a taurine-containing diet or a normal diet and were sacrificed at day 5. The AR42J pancreatic acinar cell line was incubated with/without DBTC with taurine chloramines. Apoptosis was determined by using terminal deoxynucleotidyl transferase-mediated dUTP-digoxigenin nick end labeling (TUNEL) assay. The expression of Bad and Bcl-2 proteins in the AR42J cells lysates was detected by Western blot analysis. The apoptotic index of pancreatic acinar cells in DBTC-administered rats was significantly increased. Taurine treatment inhibited pancreatic fibrosis and apoptosis of acinar cells induced by DBTC. The number of TUNEL-positive cells in the AR42J pancreatic acinar cell lines was significantly increased by the addition of DBTC. Incubation with taurine chloramines ameliorated these changes. In conclusion, taurine inhibits apoptosis of pancreatic acinar cells and pancreatitis in experimental chronic pancreatitis.

New large bowel segmentation on plain abdominal radiography in comparison with the conventional method.

Plain abdominal radiography is a very basic examination and plays an important role in primary care. The objectives of this study were to clarify colon distributions on plain abdominal radiographs. Forty-three healthy volunteers underwent gastric fluoroscopy, and 2 hours later, plain abdominal radiography in the supine position. A region of interest (ROI) was defined uniformly on each X-ray image to divide the image into 600 zones. The area corresponding to the large bowel within the ROI was divided into 4 segments (ascending colon, transverse colon, descending colon, and sigmoid colon + rectum). The percentage of barium in each segment relative to the total volume of barium used was calculated to evaluate the percent ROI occupancy. The large bowel covered 76.7% of the entire ROI, with the percent duplication being 55%. The duplicated area corresponded to the transverse colon region. When the method proposed by Arhan et al. was used, the percentage of the colon actually present in each segment relative to that determined theoretically was 99.6% for the right colon segment, 92.2& for the left colon segment, and 92.2% for the sigmoid/rectal segment. However, in cases in which the transverse colon descended partially from the fifth lumbar vertebra, the percentage occupied by the sigmoid colon + rectum decreased to 57.2%. We applied a new large bowel segmentation method especially for patients with ptosis, by devising a line joining the lateral side of the right lesser pelvis and the lower ends of both sacroiliac joints.

Measurement and correlates of empathy among female Japanese physicians.

BACKGROUND: The measurement of empathy is important in the assessment of physician competence and patient outcomes. The prevailing view is that female physicians have higher empathy scores compared with male physicians. In Japan, the number of female physicians has increased rapidly in the past ten years. In this study, we focused on female Japanese physicians and addressed factors that were associated with their empathic engagement in patient care. METHODS: The Jefferson Scale of Empathy (JSE) was translated into Japanese by using the back-translation procedure, and was administered to 285 female Japanese physicians. We designed this study to examine the psychometrics of the JSE and group differences among female Japanese physicians. RESULTS: The item-total score correlations of the JSE were all positive and statistically significant, ranging from .20 to .54, with a median of .41. The Cronbach's coefficient alpha was .81. Female physicians who were practicing in "people-oriented" specialties obtained a significantly higher mean empathy score than their counterparts in "procedure-" or "technology-oriented" specialties. In addition, physicians who reported living with their parents in an extended family or living close to their parents, scored higher on the JSE than those who were living alone or in a nuclear family. CONCLUSIONS: Our results provide support for the measurement property and reliability of the JSE in a sample of female Japanese physicians. The observed group differences associated with specialties and living arrangement may have implications for sustaining empathy. In addition, recognizing these factors that reinforce physicians' empathy may help physicians to avoid career burnout.

[Medical support on human resources and clinical laboratory in Myanmar].

I have been involved in medical cooperation programs between Myanmar and Japan for over 10 years. The purpose of the first visit to Myanmar was the investigation of hepatitis C spreading among thalassemia patients. I learned that the medical system was underdeveloped in this country, and have initiated several cooperation programs together with Professor Shigeru Okada, such as the "Protection against hepatitis C in Myanmar", "Scientist exchange between the Ministry of Health, Myanmar and Okayama University", and "Various activities sponsored by a Non-Profit Organization". As for clinical laboratories, the laboratory system itself is pre-constructed and the benefit of a clinical laboratory in modern medicine is not given to patients in Myanmar. The donation of drugs and reagents for laboratory tests is helpful, but it will be more helpful to assist the future leaders to learn modern medicine and develop their own various systems to support modern medicine. Our activity in the cooperation program is described.

Long-term management of hepatitis C-seropositive subjects with AntiOxidant Biofactor (AOB), a fermented food supplement.

The efficacy of AntiOxidant Biofactor (AOB) for the management of apparently healthy subjects with chronic hepatitis C infection was investigated. A total of 60 subjects (35 males, 25 females) participated in the trial. AOB was given orally in 2 packs (3 g per pack) 3 times per day. 17 subjects had taken AOB for 3 years, 31 subjects up to 2 years, and 41 subjects up to one year. The initial mean (SD) serum alamine aminotransferase (ALT) level was 46.3+/-35.4 IU/L, and significant (p0.05, paired t-test) reductions in the mean serum ALT levels were observed at 6 months (38.6+/-21.5 IU/L), 18 months (31.9+/-18.1 IU/L), 2 years (31.2+/-14.6 IU/L), and 3 years (28.0+/-15.9 IU/L). Those presenting with high serum ALT levels (30 subjects) demonstrated significant levels (p0.05, paired t-test) of reduction in the mean serum ALT levels at 6, 12, 18, 24, and 36 months of treatment. No side effects were observed and the AOB treatment was well tolerated by all subjects.

Prevalence and factors associated with hepatitis C virus infection among Myanmar blood donors.

We studied the prevalence of hepatitis C virus infection among blood donors from 3 hospitals of Central Myanmar and 7 hospitals of Lower Myanmar in the Yangon area, and analyzed the factors associated with the infection. The study period was from November, 2005 to June, 2007. A pre-tested questionnaire was used to obtain information on age, ethnic group, marital status, tattooing, body piercing, history of receiving transfusions, and liver diseases in self and in sexual partners. Data on seropositivity to hepatitis C, hepatitis B and human immunodeficiency virus infections were recorded. A total of 65,240 blood donors participated in the study. Their ages ranged from 18 years to 60 years (mean±SD=29.5±9.3). The male-to-female ratio was 6:1. The prevalence of the antibody to hepatitis C was found to be 0.95% with varying rates (0.34 to 2.03) among hospitals. Females had a slightly higher rate (1.06%) than males (0.93%) (p=0.237). Multivariate analyses revealed the following factors to be related to HCV infection:HIV infection, odds ratio (OR)=3.0 (p=0.003); history of liver disease, OR=8.9 (p=0.001);and age 30 years and above, OR=2.6 (p=0.001). We discuss the varying prevalences of HCV around the world.

Prediction of number of apheresis procedures necessary in healthy donors to attain minimally required peripheral blood CD34+ cells.

BACKGROUND: Allogeneic peripheral blood stem cell (PBSC) transplantation is widely performed as a curative therapy for hematopoietic malignancies. Donors for PBSC harvest (PBSCH) are usually healthy subjects and undergo granulocyte-colony-stimulating factor treatment and apheresis procedures. A considerable proportion of donors experience poor mobilization, necessitating additional harvesting or marrow collection or remobilization. Although some characteristics have been reported to correlate with poor mobilization, they may not be taken into account in selecting PBSC donors. To protect healthy donors, it is preferable to predict the number of apheresis procedures needed for PBSCH before the procedure is initiated. STUDY DESIGN AND METHODS: A retrospective cohort study of 83 subjects was conducted, using statistical models to predict the probability of obtaining a sufficient number of CD34+ cells (>or=2.0 x 10(6)/kg) in the first to the third apheresis procedures and the probability of failure to obtain sufficient cells within three apheresis sessions. This study explored potential candidate factors in an ordinal probit regression analysis. RESULTS: Significant factors predicting successful PBSCH were donor age, donor sex, and body weight difference between donor and recipient. The predictive model showed good agreement with the observed number of apheresis sessions. Simulation tables are presented with this model. CONCLUSION: The statistical model developed to predict the number of apheresis procedures for PBSCH may be useful for planning PBSCH in clinical practice.

[Effect of ISO15189 accreditation on Okayama University Hospital].

The evaluation of medical laboratories by a third person has recently become popular in Japan. Establishing a quality management system, putting it into practice, and maintaining and improving it are considered to achieve the standardization, efficacy improvement, and safety management of laboratory activities. The Departments of Clinical Laboratory and Blood Transfusion, Okayama University Hospital was awarded ISO15189 accreditation on July 6, 2007. In this paper, we describe the process of ISO15189 accreditation in our institute. As an outcome of ISO accreditation, the frequency of laboratory incidents was reduced through seeking the causes of incidents and performing continual improvement according to ISO 15189: "Plan, Do, Check, Action". Additionally the motivation and competence of laboratory staff concerned have also improved, consequently leading to a favorable influence on other laboratory staff members. Therefore, we conclude that ISO15189 is an effective tool for medical laboratory management.

Hepatitis B virus DNA in liver tissue and risk for hepatocarcinogenesis in patients with hepatitis C virus-related chronic liver disease. A prospective study.

AIMS: To prospectively study whether occult hepatitis B virus (HBV) infection can promote the development of hepatocellular carcinoma (HCC) in patients with hepatitis C virus (HCV)-related chronic liver disease. In addition, to evaluate the difference among HBV DNA-negative patients and patients with high and low HBV copy numbers. METHODS: A total of 167 patients with HCV-related chronic liver disease without HBV surface antigen (HBsAg) were studied. HBV DNA in liver tissue was determined using polymerase chain reaction (PCR). RESULTS: HBV DNA was detected in 9 of 167 patients (5.4%) by single PCR and in 25 patients (15.0%) by nested PCR. HCC developed in 12 of 167 patients (7.2%). Ten of 142 HBV DNA-negative patients (7.0%) and 2 of 9 patients with a high HBV copy number (22.2%) developed HCC, whereas none of 16 patients with a low HBV copy number developed HCC. The incidence rate of HCC in patients with a high HBV copy number was significantly higher than in HBV DNA-negative patients and patients with low HBV copy number. CONCLUSION: A high amount of HBV DNA in liver tissue of HBsAg-negative patients with HCV-related liver disease might be associated with HCC development.

Oral administration of taurine improves experimental pancreatic fibrosis.

BACKGROUND AND AIM: The mechanism of pancreatic fibrosis is unclear. Taurine is used in the clinical treatment of a wide variety of diseases, but its effect on improving pancreatic fibrosis is unknown. We examined whether a diet with added taurine improves pancreatic fibrosis induced by dibutyltin dichloride (DBTC) in an experimental chronic pancreatitis rat model. In addition, we examined the influence of taurine on pancreatic stellate cells. METHODS: Pancreatic fibrosis was induced by DBTC. Rats were fed a taurine-containing diet or a normal diet and were killed at 4 weeks. Pancreatic stellate cells were isolated from male Wistar rats. Cultured pancreatic stellate cells were incubated with or without taurine chloramine. Type I collagen and transforming growth factor-beta1 secretion was evaluated by ELISA, and matrix metalloproteinase activity was assessed by gelatin zymography. Interleukin-6, interleukin-2, and transforming growth factor-beta1 levels in the supernatants of pancreatic tissue homogenates were measured. RESULTS: Pancreatic fibrosis induced by DBTC was improved remarkably by the oral administration of the taurine-containing diet. Taurine chloramine decreased type I collagen, transforming growth factor-beta1, and matrix metalloproteinases 2 of the pancreatic stellate cell culture supernatant. Increased interleukin-6 and decreased interleukin-2 were found in the supernatants of the pancreatic tissue homogenates of DBTC-induced pancreatitis rats compared with other groups. CONCLUSION: The oral administration of taurine improves pancreatic fibrosis. Taurine chloramine inhibits transforming growth factor-beta1 produced from activated pancreatic stellate cells and improves pancreatic fibrosis.

Hepatitis C virus genotype distribution in Myanmar: Predominance of genotype 6 and existence of new genotype 6 subtype.

AIM: This study was performed to determine the prevalence and distribution of hepatitis C virus (HCV) genotypes in Myanmar. METHODS: A total of 1333 peripheral blood samples were collected from four different border cities of Myanmar. The anti-HCV antibody-positive serum samples were identified. HCV was genotyped by reverse transcriptase polymerase chain reaction, direct DNA sequencing and phylogenetic analysis on the partial core genome. RESULTS: The overall prevalence of HCV infection was 11.6% (154/1333). Regionally, it was 13.5% (47/349) in the north-eastern city, 12.8% (64/501) in the north-western city, 4.2% (16/380) in the southern city and 26.2% (27/103) in the western city. HCV was genotyped in 145/154 (94.2%) samples. Genotype 6 was the most prevalent genotype in this study (71/145, 49%), followed by genotype 3 (57/145, 39.3%), genotype 1 (16/145, 11%), and genotype 2 (1/145, 0.7%). Genotype 6 was mostly found in the northern cities and genotype 3 in the southern and western cities of Myanmar. Multiple HCV genotypes/subtypes were successfully characterized as 1a, 1b, 2a, 3a, 3b, 6m, 6n, and a new 6 subtype. Among them, subtype 6n was the most predominant subtype (38.6%), followed by subtype 3b (29.7%), 3a (9.6%), 6m (9%), 1b (6.9%), 1a (4.1%), new 6 subtype (1.4%) and 2a (0.7%). Subtype 6n was more widely distributed in the northern cities whereas subtype 3b was more common in the western city. The newly discovered genotype 6 subtype was from the northern cities. CONCLUSIONS: The results indicate there are regional differences of HCV genotype distribution in Myanmar. There is a distinct geographic variation from other South-East Asian countries in terms of the existence of the new genotype 6 subtype.

[Fibrosis markers in heavy alcohol drinkers].

Alcoholic intake has increased in society in recent years. gamma-GTP is used as a marker of liver damage by alcohol intake, but there is no reliable marker of pancreatic fibrosis. We used animal experiments and clinical data to identify a new reliable marker of early-stage pancreatic fibrosis. Pancreatic fibrosis is induced by intra-peritoneal injection of diethyldithiocarbamate. Pancreas tissue was extracted and measured. Human pure pancreatic juice was collected by endoscopic procedures. Prolyl hydroxylase in pancreas tissue is increased in the early stage of pancreatic fibrosis. Secretion of matrix metalloproteinase from pancreatic stellate cells is increased by diethyldithiocarbamate stimulation. Pancreatic stellate cells, prolyl hydroxylase and a tissue inhibitor of metalloproteinase in human pure pancreatic juice is increased in heavy alcohol drinkers and normalized in former alcohol drinkers. Active matrix metalloproteinase 2 is detected in pure pancreatic juice of chronic pancreatitis patients. Treatment with oral camostat increases pancreatic secretory trypsin inhibitor in chronic pancreatitis patients. Experimental and clinical data indicated that matrix metalloproteinase 2 and prolyl hydroxylase are candidates as markers of early-stage pancreatic fibrosis. Clinical data showed that tissue inhibitor of metalloproteinase and pancreatic secretory trypsin inhibitor in pure pancreatic juice had potential as markers of early-stage pancreatic fibrosis.

Three type 6 hepatitis C virus subgroups among blood donors in the Yangon area of Myanmar are identified as subtypes 6m and 6n, and a novel subtype by sequence analysis of the core region.

Previously, using phylogenetic analysis of NS5b sequences, we found that three type 6 variant subgroups (M6-1, M6-2 and M6-3) exist in Myanmar. According to the new nomenclature of hepatitis C, M6-1 and M6-2 belong to subtypes 6m and 6n, respectively, but M6-3 is unassigned. In this study, we sequenced and phylogenetically analyzed the core region of these type 6 variant subgroups. Serum samples assigned as 6m or 6n by NS5b sequence were also identified as 6 m or 6n by core region analysis. The M6-3 (sample name MYAN-3E-3) remained unassigned to a subgroup based on its core region analysis. The findings of this study suggest that either the core region or the NS5b region can be analyzed for HCV subtype classification.

[Effort in standardization of control survey using pooled fresh serum in Okayama Prefecture].

Control survey of the clinical laboratories in Okayama Prefecture has been continued once a year for 18 years using commercially available control serum. The purpose of our control survey is to minimize the difference of laboratory test data among laboratories. It is clear, however, that one time a year survey leads to miss-evaluation of the difference in laboratory test result. Furthermore the data difference caused by difference of assay methods, for example dry chemistry and wet chemistry, is not properly evaluated by control survey using control serum, since test result is affected by matrix effect. Therefore we have developed a monthly control survey program, Okayama cross-check program, using pooled fresh serum among laboratories of middle-large scale hospitals and commercial laboratories in Okayama prefecture. In this paper we describe the details of Okayama cross-check program and the usefulness of the program to reduce the difference among laboratory facilities. This result was attained by cooperation of the Okayama prefectural medical association, Okayama association of medical technologists, and commercial laboratories in Okayama.

Functional promoter upstream p53 regulatory sequence of IGFBP3 that is silenced by tumor specific methylation.

BACKGROUND: Insulin-like growth factor binding protein (IGFBP)-3 functions as a carrier of insulin-like growth factors (IGFs) in circulation and a mediator of the growth suppression signal in cells. There are two reported p53 regulatory regions in the IGFBP3 gene; one upstream of the promoter and one intronic. We previously reported a hot spot of promoter hypermethylation of IGFBP-3 in human hepatocellular carcinomas and derivative cell lines. As the hot spot locates at the putative upstream p53 consensus sequences, these p53 consensus sequences are really functional is a question to be answered. METHODS: In this study, we examined the p53 consensus sequences upstream of the IGFBP-3 promoter for the p53 induced expression of IGFBP-3. Deletion, mutagenesis, and methylation constructs of IGFBP-3 promoter were assessed in the human hepatoblastoma cell line HepG2 for promoter activity. RESULTS: Deletions and mutations of these sequences completely abolished the expression of IGFBP-3 in the presence of p53 overexpression. In vitro methylation of these p53 consensus sequences also suppressed IGFBP-3 expression. In contrast, the expression of IGFBP-3 was not affected in the absence of p53 overexpression. Further, we observed by electrophoresis mobility shift assay that p53 binding to the promoter region was diminished when methylated. CONCLUSION: From these observations, we conclude that four out of eleven p53 consensus sequences upstream of the IGFBP-3 promoter are essential for the p53 induced expression of IGFBP-3, and hypermethylation of these sequences selectively suppresses p53 induced IGFBP-3 expression in HepG2 cells.

Treatment of elderly patients with early gastric cancer by endoscopic submucosal dissection using an insulated-tip diathermic knife.

OBJECTIVE: In recent years, the number of elderly patients with early gastric cancer (EGC) has steadily been increasing. In our institute, endoscopic submucosal dissection (ESD) involving the use of an insulated-tip diathermic knife (IT-ESD) was introduced for the treatment of mucosal gastric carcinoma in 1996. The purpose of this study was to evaluate the effectiveness of IT-ESD for the treatment of elderly patients with EGC. MATERIALS AND METHODS: A total of 144 patients with EGC were treated at Shikoku Cancer Center in the 5-year period from January 2000 to December 2004, including 53 patients over 75 years old. The performance status (PS) for all patients was less than 2. We compared patient's backgrounds, the one-piece resection rate, complete resection (CR) rate, operation time, bleeding rate, perforation rate, blood pressure, and peripheral oxygen saturation (SpO(2)) between patients over 75 years of age (elderly group) and the remaining 91 younger patients (non-elderly group). RESULTS: The rate of having underlying disease was significantly higher for the elderly group (p<0.05) and we found no significant difference for the one-piece resection rate, CR rate, operation time, bleeding rate, and perforation rate between the 2 groups. There were also no significant differences for the frequency of the use of oxygen, pressor and depressor between the 2 groups. CONCLUSION: There was no significant difference in the outcome resulting from ESD between the 2 groups. Our study proves that ESD is a feasible treatment for elderly patients with EGC PS of less than 2.

Suppressed erythropoiesis after ABO-minor-mismatched living-donor lobar lung transplantation.

Anti-A and/or anti-B antibodies synthesized by lymphoid tissues contained in ABO-mismatched solid organ grafts have been reported to cause hemolytic anemia, but not suppressed red blood cell production. A case of living-donor lobar lung transplantation is presented in which suppression of erythropoiesis was associated with ABO mismatched organ transplantation.

Matrix metalloproteinase-2 in pancreatic juice for diagnosis of pancreatic cancer.

INTRODUCTION: Matrix metalloproteinase-2 (MMP-2) has an activity to degrade type IV collagen and is associated with invasion angiogenesis of malignant tumor. AIM: A diagnostic value of MMP-2 in pancreatic juice was studied in the diagnosis of pancreatic cancer. METHODOLOGY: Using gelatin zymography, active MMP-2 and proMMP-2 were determined in pancreatic juice obtained endoscopically from 12 patients with pancreatic cancer, 11 with chronic pancreatitis, and 7 control subjects. RESULTS: ProMMP-2 was detected in 12 of 12 patients (100%) with pancreatic cancer, 6 of 11 (54.5%) with chronic pancreatitis, and 3 of 7 (42.9%) controls. Active MMP-2 was detected in 11 patients (91.6%) with pancreatic cancer, 2 (18.2%) with chronic pancreatitis, and none of the control subjects. An activation ratio of MMP-2 (active MMP-2/total MMP-2) in pancreatic juice is significantly higher in pancreatic cancer (23.4 +/- 4.4%, mean +/- SE) than in chronic pancreatitis (2.1 +/- 1.7%) and controls (0%) (p < 0.01). Active MMP-2 was also detected in pancreatic juice from three cases of small pancreatic cancer (tumor <2 cm in diameter). CONCLUSION: Our observation suggests that detection of active MMP-2 in pancreatic juice using gelatin zymography may be useful for the diagnosis of pancreatic cancer.

Microencapsulated multicellular spheroid of rat hepatocytes transplanted intraperitoneally after 90% hepatectomy.

BACKGROUND/AIMS: The aim of this preliminary study was to investigate the metabolic activity of hepatocyte spheroids encapsulated with agarosehydrogel in vitro and the effect of encapsulated hepatocyte spheroids transplanted intraperitoneally on survival in the 90% hepatectomized rats for preparing future xeno-hepatocyte spheroids transplantation. METHODOLOGY: Hepatocytes were prepared by Seglen's method and cultured with epidermal growth factor. Hepatocytes formed spheroids 4 days after inoculation. Hepatocyte spheroids were encapsulated into 5% concentration agarose-hydrogel microbeads before being transplanted into the peritoneum of the rats. RESULTS: Hepatocyte spherois showed better metabolic activity compared to isolated hepatocytes. Hepatocyte spheroids function cultured in vitro were not deteriorated by encapsulation with agarose-hydrogel. Survival rates of 90% hepatectomized rats were improved by the intraperitoneal transplantation of encapsulated hepatocyte spheroids. CONCLUSIONS: Rat hepatocyte spheroids encapsulated with agarose-hydrogel and transplanted intraperitoneally were considered as effective for the improvement of survival rates in acute liver failure produced by 90% hepatectomy.